Identification of miR-21 targets in breast cancer cells using a quantitative proteomic approach

Yi Yang, Raghothama Chaerkady, Michael A. Beer, Joshua T. Mendell, Akhilesh Pandey

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

MicroRNA (miRNA) play essential roles in biological processes ranging from cellular proliferation to apoptosis. Recently, miRNA have also been implicated in a number of diseases including cancers. However, the targets of most miRNA remain unknown. The majority of reports describing identification of miRNA targets are based on computational approaches or detection of altered mRNA levels despite the fact that most miRNA are thought to regulate their targets primarily at the level of translational inhibition in animals. miR-21 is a miRNA with oncogenic activity that is involved in various cancer-related processes such as invasion and migration. Given the importance of miR-21 in tumorigenesis, we employed a quantitative proteomic strategy to systematically identify potential targets of miR-21. By knocking down the expression of endogenous miR-21 in MCF-7 breast cancer cells, we observed an increase in the abundance of 58 proteins, implying that they could be potential targets of miR-21. Validation of 12 of these candidate targets in luciferase assays showed that 6 of them were likely direct targets of miR-21. Importantly, the mRNA of the majority of the candidate targets tested did not show a concomitant increase in abundance. Overall, our results demonstrate that miR-21 affects the expression of many of its targets through translational inhibition and highlights the utility of proteomic approaches for identifying miRNA targets.

Original languageEnglish (US)
Pages (from-to)1374-1384
Number of pages11
JournalProteomics
Volume9
Issue number5
DOIs
StatePublished - Mar 2009

Fingerprint

MicroRNAs
Proteomics
Cells
Breast Neoplasms
Biological Phenomena
Messenger RNA
Luciferases
Assays
Neoplasms
Carcinogenesis
Animals
Cell Proliferation
Apoptosis
Proteins

Keywords

  • Breast cancer cells
  • MicroRNA
  • Quantitative analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

Cite this

Identification of miR-21 targets in breast cancer cells using a quantitative proteomic approach. / Yang, Yi; Chaerkady, Raghothama; Beer, Michael A.; Mendell, Joshua T.; Pandey, Akhilesh.

In: Proteomics, Vol. 9, No. 5, 03.2009, p. 1374-1384.

Research output: Contribution to journalArticle

Yang, Yi ; Chaerkady, Raghothama ; Beer, Michael A. ; Mendell, Joshua T. ; Pandey, Akhilesh. / Identification of miR-21 targets in breast cancer cells using a quantitative proteomic approach. In: Proteomics. 2009 ; Vol. 9, No. 5. pp. 1374-1384.
@article{99101c3f2fc447b593486c007a235b07,
title = "Identification of miR-21 targets in breast cancer cells using a quantitative proteomic approach",
abstract = "MicroRNA (miRNA) play essential roles in biological processes ranging from cellular proliferation to apoptosis. Recently, miRNA have also been implicated in a number of diseases including cancers. However, the targets of most miRNA remain unknown. The majority of reports describing identification of miRNA targets are based on computational approaches or detection of altered mRNA levels despite the fact that most miRNA are thought to regulate their targets primarily at the level of translational inhibition in animals. miR-21 is a miRNA with oncogenic activity that is involved in various cancer-related processes such as invasion and migration. Given the importance of miR-21 in tumorigenesis, we employed a quantitative proteomic strategy to systematically identify potential targets of miR-21. By knocking down the expression of endogenous miR-21 in MCF-7 breast cancer cells, we observed an increase in the abundance of 58 proteins, implying that they could be potential targets of miR-21. Validation of 12 of these candidate targets in luciferase assays showed that 6 of them were likely direct targets of miR-21. Importantly, the mRNA of the majority of the candidate targets tested did not show a concomitant increase in abundance. Overall, our results demonstrate that miR-21 affects the expression of many of its targets through translational inhibition and highlights the utility of proteomic approaches for identifying miRNA targets.",
keywords = "Breast cancer cells, MicroRNA, Quantitative analysis",
author = "Yi Yang and Raghothama Chaerkady and Beer, {Michael A.} and Mendell, {Joshua T.} and Akhilesh Pandey",
year = "2009",
month = "3",
doi = "10.1002/pmic.200800551",
language = "English (US)",
volume = "9",
pages = "1374--1384",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "5",

}

TY - JOUR

T1 - Identification of miR-21 targets in breast cancer cells using a quantitative proteomic approach

AU - Yang, Yi

AU - Chaerkady, Raghothama

AU - Beer, Michael A.

AU - Mendell, Joshua T.

AU - Pandey, Akhilesh

PY - 2009/3

Y1 - 2009/3

N2 - MicroRNA (miRNA) play essential roles in biological processes ranging from cellular proliferation to apoptosis. Recently, miRNA have also been implicated in a number of diseases including cancers. However, the targets of most miRNA remain unknown. The majority of reports describing identification of miRNA targets are based on computational approaches or detection of altered mRNA levels despite the fact that most miRNA are thought to regulate their targets primarily at the level of translational inhibition in animals. miR-21 is a miRNA with oncogenic activity that is involved in various cancer-related processes such as invasion and migration. Given the importance of miR-21 in tumorigenesis, we employed a quantitative proteomic strategy to systematically identify potential targets of miR-21. By knocking down the expression of endogenous miR-21 in MCF-7 breast cancer cells, we observed an increase in the abundance of 58 proteins, implying that they could be potential targets of miR-21. Validation of 12 of these candidate targets in luciferase assays showed that 6 of them were likely direct targets of miR-21. Importantly, the mRNA of the majority of the candidate targets tested did not show a concomitant increase in abundance. Overall, our results demonstrate that miR-21 affects the expression of many of its targets through translational inhibition and highlights the utility of proteomic approaches for identifying miRNA targets.

AB - MicroRNA (miRNA) play essential roles in biological processes ranging from cellular proliferation to apoptosis. Recently, miRNA have also been implicated in a number of diseases including cancers. However, the targets of most miRNA remain unknown. The majority of reports describing identification of miRNA targets are based on computational approaches or detection of altered mRNA levels despite the fact that most miRNA are thought to regulate their targets primarily at the level of translational inhibition in animals. miR-21 is a miRNA with oncogenic activity that is involved in various cancer-related processes such as invasion and migration. Given the importance of miR-21 in tumorigenesis, we employed a quantitative proteomic strategy to systematically identify potential targets of miR-21. By knocking down the expression of endogenous miR-21 in MCF-7 breast cancer cells, we observed an increase in the abundance of 58 proteins, implying that they could be potential targets of miR-21. Validation of 12 of these candidate targets in luciferase assays showed that 6 of them were likely direct targets of miR-21. Importantly, the mRNA of the majority of the candidate targets tested did not show a concomitant increase in abundance. Overall, our results demonstrate that miR-21 affects the expression of many of its targets through translational inhibition and highlights the utility of proteomic approaches for identifying miRNA targets.

KW - Breast cancer cells

KW - MicroRNA

KW - Quantitative analysis

UR - http://www.scopus.com/inward/record.url?scp=63049092022&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=63049092022&partnerID=8YFLogxK

U2 - 10.1002/pmic.200800551

DO - 10.1002/pmic.200800551

M3 - Article

C2 - 19253296

AN - SCOPUS:63049092022

VL - 9

SP - 1374

EP - 1384

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 5

ER -